奶牛早孕因子重组蛋白的原核表达、纯化及多克隆抗体的制备

doi:10.16431/j.cnki.1671-7236.2015.04.016

Abstract

Abstract: The study was aimed to express and purify the dairy cattle early pregnancy factor (EPF) protein and prepare EPF polyclonal antibody. The recombinant plasmid pET32a-EPF was expressed in E.coli BL21 (DE3). Polyclonal antibodies were developed by immunizing BALB/c mice with the SDS-PAGE gel extraction purified fusion protein. The recombinant protein and the polyclonal antibody were separately detected by Western blotting and ELISA. The results showed that the recombinant protein with a molecular weight of 37 ku was expressed successfully in E.coli and its content was approximately 48.6% of total bacteria proteins. The purity of target protein could reach 93% after purification. The recombinant protein was recognized by the prepared polyclonal antibody in Western blotting analysis. The antibody titer was 1:12 800.These results indicated that we had successfully obtained and purified the dairy cattle EPF protein, which laid a foundation for the further study of dairy cattle pregnancy diagnosis.

Key words: early pregnancy factor; prokaryotic expression; SDS-PAGE; Western blotting; ELISA

CLC Number:

S823

LIU Yun, JIA Bin, SHI Feng, LI Xin, LI Hong-tao, ZHANG Yong-sheng, NING Meng-ying, JI Jun-ming, QIN Bing-yan. Prokaryotic Expression,Purification of Early Pregnancy Factor Protein of the Dairy Cattle and Preparation of Polyclonal Antibody against it[J]. , 2015, 42(4): 877-882.

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Prokaryotic Expression,Purification of Early Pregnancy Factor Protein of the Dairy Cattle and Preparation of Polyclonal Antibody against it

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